LOCUS       Exported               13621 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDART, complete sequence.
ACCESSION   KP126802
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13621)
  AUTHORS   Ryu J, Lee U, Park J, Yoo DH, Ahn JH.
  TITLE     A Vector System for ABC Transporter-Mediated Secretion and 
            Purification of Recombinant Proteins in Pseudomonas Species
  JOURNAL   Appl. Environ. Microbiol. 81 (5), 1744-1753 (2015)
  PUBMED    25548043
REFERENCE   2  (bases 1 to 13621)
  AUTHORS   Ryu J, Lee U, Park J, Yoo D-H., Ahn JH.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-NOV-2014) Dept. Biology, Korea Science Academy of 
            KAIST, #899, Tanggam 3-Dong, Busanjin-Gu, Busan 614-822, Korea
REFERENCE   3  (bases 1 to 13621)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..13621
                     /organism="cloning vector pDART"
                     /lab_host="Pseudomonas fluorescens SIK W1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1591410"
     primer_bind     complement(1..17)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1002..1817
                     /codon_start=1
                     /gene="aph(3)-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2631..3025
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     oriT            3367..3454
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             4693..5664
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             6175..7014
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     protein_bind    8516..8537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8552..8582
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8590..8606
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8614..8630
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             8674..10410
                     /codon_start=1
                     /product="TliD"
                     /label=TliD
                     /note="ABC protein"
                     /protein_id="AJC64569.1"
                     /translation="MSFGSRKINHYGQAYCRGAFIQGVGEYKSILISVGCFTALINLLM
                     LVPSIYMLQVYDRVLSSQNETTLVMLTLMVVGFFAFIGTLEVIRSFIVIRIGSQLERRF
                     NLRVYKAAFERNLQRGQGHAGQALGDLTLLRQFITGPALFAFFDAPWFPLYLLVIFLFN
                     VWLGVLATAGAVLLIGLACLNEYLTKKPLGEAGAYSQQSSQLATSHLHNAETIQAMGML
                     GALRKRWFAVHSQFLGLQNTASDTGSVITSLSKTLRLCLQSLVLGLGALLVIKGDMTAG
                     MMIAGSILMGRVLSPIDQLIAVWKQWSSAKLAYQRLDDLLREFPPDSEPMKLPAPHGQV
                     SFEQVSAGPPGRRTPTLHQVSFTLGAGEVLGVLGASGSGKSTLARVLVGVWPTLGGTVR
                     LDGADIHRWDREDLGPHIGYLPQDIELFSGSIADNIARFRQADPALVVQAAQQAGVHEL
                     ILRLPHGYDTLLGDNGGGLSGGQKQRVALARALYGGPRLIVLDEPNSNLDTVGEAALAS
                     AIVQMKAQGSSVVLVTHRSSALAQADKLLVLNEGRLQRLARARRCCAHCPASRKHRRKG
                     PV"
     CDS             10451..11752
                     /codon_start=1
                     /product="TliE"
                     /label=TliE
                     /note="membrane fusion protein"
                     /protein_id="AJC64570.1"
                     /translation="MSSLTFEQRDARFFVRMGWLLTVVGAGGFFLWASLAPLDQGIPVQ
                     GTVVVSGKRKAVQTFSPGVVSRILVREGETVKQGQPLFRLDQTQNQADVQSLQAQYRLA
                     WASVARWQSERDNRSSIHFPAELSSNPDPALALVLEGQRQLFSSRREAFAREQAGIRAN
                     IDGATAQLGGMRRARSDLTAQAQSLRDQLSNLQPLADNGYIPRNRLMEYQRQLSQVQQD
                     LAQNTGESGRVEQGILESRLKLQQHSEEYQKEVRSQLADAQLRSLTLEQQLTSAGFDLQ
                     HSEINAPADGIAVNLGVHTEGAVVRAGETLLEIVPQGTRLEVEGHLPVHLVDKVGTHLP
                     VDILFTAFNQSRTPRVPGEVSLISADQMLDEKTGAPYYVLRTTLSDAAEQKLQGLVIKP
                     GMPAEMFVRTGERSLLNYLFKPLLDRAGSALTEE"
     CDS             11755..13200
                     /codon_start=1
                     /product="TliF"
                     /label=TliF
                     /note="outer membrane protein"
                     /protein_id="AJC64571.1"
                     /translation="MRSLLIAVLFSCASAHAAMGPFDVYEQALRNDPVFLGAIKERDAG
                     LENRTIGRAGLLPKLSYNYNKGRNNSQATLPDGRGGNYHDDRNYNSYGSTFSLQQPLFD
                     YEAYANYRKGVAQALFADESFRDKSQALLVRVLTYYTQALFAQDQIDIARAKKKAFEQQ
                     FQQNRHLFEQGEGTRTDILEAESRYELATAEEIEALDEQDAALRELGALIGVQSVNIDD
                     LAPLSPGFAAFSLSPANYDTWHELAISNNPTLASQRQALEVARYEVERNRAGHLPKVTA
                     YASSRQQESDSGNTYNQRYDTNTIGVEVSLPLYAGGGVSASTRQASRAMEQAEYELEGK
                     TRETLIELRRQFSACLSGVSKLRAYQKALTSAEALVVSTRQSILGGERVNLDALNAEQQ
                     LYSTRRDLAQARYDYLMAWTKLHYYAGNLRDTDLAKVDEAFGTKRAEPPAADKPPVANK
                     PPEASRPPVASGLAPPWAAQQPQ"
     regulatory      11803..11812
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      13234..13239
                     /regulatory_class="ribosome_binding_site"
     CDS             13247..13606
                     /codon_start=1
                     /gene="LARD3"
                     /product="LARD3"
                     /label=LARD3
                     /note="C-terminal signal sequence for secretion derived 
                     from TliA"
                     /protein_id="AJC64572.1"
                     /translation="MSRHMGTASELIEGRGSDGNDLIQGGKGADFIEGGKGNDTIRDNS
                     GHNTFLFSGHFGQDRIIGYQPTDRLVFQGADGSTDLRDHAKAVGADTVLSFGADSVTLV
                     GVGLGGLWSEGVLIS"
     gene            13247..13606
                     /gene="LARD3"
                     /label=LARD3
     misc_feature    13250..13279
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    13280..13291
                     /label=factor Xa cleavage site
                     /note="factor Xa cleavage site"
     CDS             13280..13291
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"